Abstract
Attenuated Salmonella can invade tumor cells and acts as a eukaryotic expression vector for gene propagation. We constructed a bi-gene, eukaryotic co-expression DNA vaccine of Mycobacterium tuberculosis heat shock protein 70 (mtHSP70) and Herpes simplex virus-thymidine kinase (HSV-tk) and used attenuated Salmonella as a vector to treat murine melanoma. In vitro, recombinant Salmonella can carry plasmid stably and can invade into the cytoplasm of B16 tumor cells expressing the protein of the mtHSP70/HSV-tk gene by Western blot assay. In vivo, after the recombinant Salmonella was injected into tumors, the HSV-tk precursor drug ganciclovir (GCV) was administered to start the HSV-tk killing of tumor cells. We found that the mtHSP70/HSV-tk recombinant bacteria can raise CD8+ T lymphocytes in peripheral blood by flow cytometry and in tumor tissues by immunofluorescence detection, increase IFN‑γ contents in tumor tissue by ELISA and significantly suppress tumor growth.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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CD8-Positive T-Lymphocytes / immunology
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CD8-Positive T-Lymphocytes / metabolism
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Cancer Vaccines / genetics
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Cancer Vaccines / immunology
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Cancer Vaccines / pharmacology*
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Cell Line, Tumor
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Ganciclovir / administration & dosage
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Genetic Therapy / methods
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Genetic Vectors / genetics
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Genetic Vectors / immunology
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HSP70 Heat-Shock Proteins / genetics
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HSP70 Heat-Shock Proteins / immunology*
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Interferon-gamma / immunology
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Interferon-gamma / metabolism
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Melanoma, Experimental / microbiology
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Melanoma, Experimental / pathology
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Melanoma, Experimental / therapy*
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Mice
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Mice, Inbred C57BL
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Mycobacterium tuberculosis / genetics
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Plasmids / genetics
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Salmonella / genetics
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Salmonella / immunology*
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Salmonella Infections / genetics
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Salmonella Vaccines / genetics
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Salmonella Vaccines / immunology
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Salmonella Vaccines / pharmacology*
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Simplexvirus / enzymology*
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Simplexvirus / genetics
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Thymidine Kinase / genetics
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Thymidine Kinase / immunology*
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Transfection / methods
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Vaccines, Attenuated / genetics
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Vaccines, Attenuated / immunology
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Vaccines, Attenuated / pharmacology
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Viral Proteins / genetics
Substances
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Cancer Vaccines
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HSP70 Heat-Shock Proteins
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Salmonella Vaccines
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Vaccines, Attenuated
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Viral Proteins
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Interferon-gamma
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Thymidine Kinase
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Ganciclovir